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Functional characterization of common protein variants in the efflux transporter ABCC11 and identification of T546M as functionally damaging variant

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Abstract

Multidrug resistance protein 8 (ABCC11) is an efflux transporter for anionic lipophilic compounds, conferring resistance to antiviral and anticancer agents like 5-fluorouracil (5-FU). ABCC11 missense variants may contribute to variability in drug response but functional consequences, except for the ‘earwax variant’ c.538G>A, are unknown. Using the ‘Screen and Insert’ technology, we generated human embryonic kidney 293 cells stably expressing ABCC11 missense variants frequently occurring in different ethnic populations: c.57G>A, c.538G>A, c.950C>A, c.1637C>T, c.1942G>A, c.4032A>G. A series of in silico prediction analyses and in vitro plasma membrane vesicle uptake, immunoblotting and immunolocalization experiments were undertaken to investigate functional consequences. We identified c.1637C>T (T546M), previously associated with 5-FU-related toxicity, as a novel functionally damaging ABCC11 variant exhibiting markedly reduced transport function of 5-FdUMP, the active cytotoxic metabolite of 5-FU. Detailed analysis of 14 subpopulations revealed highest allele frequencies of c.1637C>T in Europeans and Americans (up to 11%) compared with Africans and Asians (up to 3%).

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Acknowledgements

We gratefully acknowledge Sabine Rekersbrink and Silvia Hübner for expert technical assistance. RA, TL, ES, MS and ATN were supported by the Robert Bosch Stiftung, Stuttgart, Germany, RA and TL were supported by the German Research Foundation DFG (LA 2406/2-1). GJ was supported by the German Research Foundation DFG (JE 234/4-1). ATN and MS were supported by the 7FP EU Initial Training Network program‚ FightingDrugFailure’ (GA-2009–238132). MS and ES were supported in part by the Federal Ministry for Education and Research (BMBF, Berlin, Germany; grant 03 IS 2061C and 0315755).

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Authors and Affiliations

  1. Dr Margarete Fischer-Bosch Institute of Clinical Pharmacology, Stuttgart and University of Tübingen, Stuttgart, Germany,

    R Arlanov, T Lang, E Schaeffeler, M Schwab & A T Nies

  2. Department of Pharmacology, Center of Drug Absorption and Transport (C_DAT), University Medicine of Greifswald, Greifswald, Germany

    G Jedlitschky

  3. RIKEN Center for Life Science Technologies, Yokohama, Japan

    T Ishikawa

  4. Department of Clinical Pharmacology, Institute of Experimental and Clinical Pharmacology and Toxicology, University Hospital of Tübingen, Tübingen, Germany

    M Schwab

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  1. R Arlanov
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  2. T Lang
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  3. G Jedlitschky
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Correspondence to M Schwab.

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Arlanov, R., Lang, T., Jedlitschky, G. et al. Functional characterization of common protein variants in the efflux transporter ABCC11 and identification of T546M as functionally damaging variant. Pharmacogenomics J 16, 193–201 (2016). https://doi.org/10.1038/tpj.2015.27

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